BACKGROUND-AIM Leishmaniasis is a protozoan disease caused by parasites belonging to the genus Leishmania. In the human host, these parasites invade macrophages where they develop into intracellular amastigotes and multiply within phagolysosomes. Host cells can control the infection initially through the triggering of innate immune responses. NOD-like receptors (NLRs) are a family of innate immune cytosolic receptors able to recognize pathogen-associated molecular patterns. NOD1 and NOD2 detect pathogens that are able to invade and multiply intracellularly. Once stimulated, these receptors induce the activation of NF-κB and MAPKs, which lead to the transcription of genes involved in inflammation and immune responses. The aim of this work was to evaluate the role of NOD2 in some innate immune responses of macrophages infected with L. tropica. In particular, the production of TNF-a, or nitric oxide (NO), and the expression of inducible NO synthase (iNOS) were evaluated METHODS Immortalized bone marrow-derived macrophages (BMDM) from wild type (WT) C57Bl/6 or knockout (KO) mice for NOD2 were used. The levels of TNF-a released into the supernatants of BMDM-WT or -NOD2-KO treated with L. tropica were measured by ELISA. Also, the presence of nitrite was evaluated, through the Griess test, as an indication of nitric oxide (NO) production. Finally, protein and gene expression levels of iNOS were retrieved by Western blot analysis and realtime PCR, respectively. RESULTS The involvement of NOD2 in BMDM treated with L. tropica was elucidated as the levels of TNF-a or NO released from BMDM-WT infected with L. tropica were significantly higher compared to the BMDM-NOD2-KO. On the other side, the expression of iNOS (RNA and protein) was higher in BMDM-WT treated with L. tropica than in BMDM-NOD2-KO. CONCLUSIONS Altogether, these data indicated a crucial role of NOD2 for these innate immune responses of BMDM infected with L.tropica.

Involvement of NOD2 in macrophage response to leishmania tropica infection / D. Scaccabarozzi, N. Basilico, L. Cavicchini, D. Taramelli, Y. Corbett. ((Intervento presentato al convegno Pathobiology: from molecular disease to clinical application tenutosi a Firenze nel 2019.

Involvement of NOD2 in macrophage response to leishmania tropica infection

D. Scaccabarozzi
Primo
;
N. Basilico
Secondo
;
L. Cavicchini;D. Taramelli;Y. Corbett
Ultimo
2019

Abstract

BACKGROUND-AIM Leishmaniasis is a protozoan disease caused by parasites belonging to the genus Leishmania. In the human host, these parasites invade macrophages where they develop into intracellular amastigotes and multiply within phagolysosomes. Host cells can control the infection initially through the triggering of innate immune responses. NOD-like receptors (NLRs) are a family of innate immune cytosolic receptors able to recognize pathogen-associated molecular patterns. NOD1 and NOD2 detect pathogens that are able to invade and multiply intracellularly. Once stimulated, these receptors induce the activation of NF-κB and MAPKs, which lead to the transcription of genes involved in inflammation and immune responses. The aim of this work was to evaluate the role of NOD2 in some innate immune responses of macrophages infected with L. tropica. In particular, the production of TNF-a, or nitric oxide (NO), and the expression of inducible NO synthase (iNOS) were evaluated METHODS Immortalized bone marrow-derived macrophages (BMDM) from wild type (WT) C57Bl/6 or knockout (KO) mice for NOD2 were used. The levels of TNF-a released into the supernatants of BMDM-WT or -NOD2-KO treated with L. tropica were measured by ELISA. Also, the presence of nitrite was evaluated, through the Griess test, as an indication of nitric oxide (NO) production. Finally, protein and gene expression levels of iNOS were retrieved by Western blot analysis and realtime PCR, respectively. RESULTS The involvement of NOD2 in BMDM treated with L. tropica was elucidated as the levels of TNF-a or NO released from BMDM-WT infected with L. tropica were significantly higher compared to the BMDM-NOD2-KO. On the other side, the expression of iNOS (RNA and protein) was higher in BMDM-WT treated with L. tropica than in BMDM-NOD2-KO. CONCLUSIONS Altogether, these data indicated a crucial role of NOD2 for these innate immune responses of BMDM infected with L.tropica.
13-set-2019
Settore MED/04 - Patologia Generale
Settore MED/07 - Microbiologia e Microbiologia Clinica
Settore MED/46 - Scienze Tecniche di Medicina di Laboratorio
Involvement of NOD2 in macrophage response to leishmania tropica infection / D. Scaccabarozzi, N. Basilico, L. Cavicchini, D. Taramelli, Y. Corbett. ((Intervento presentato al convegno Pathobiology: from molecular disease to clinical application tenutosi a Firenze nel 2019.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/707064
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